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. Author manuscript; available in PMC: 2014 Apr 25.
Published in final edited form as: Cell Rep. 2013 Apr 11;3(4):1140–1152. doi: 10.1016/j.celrep.2013.03.025

Figure 1. Induction and isolation of NC from Sox10::GFP hESCs using a modified dual SMAD inhibition protocol.

Figure 1

(A) A dual SMAD inhibition (DSi) protocol can be modified to support highly efficient induction of a neural crest (NC) population by optimizing BMP, TGF-β, and Wnt signaling. (B) NC conditions support induction of a Sox10::GFP expressing population that co-expresses neural crest markers HNK-1 and p75 while down-regulating ZO-1 and PAX6 expressing CNS. Scale bars represent 200µm. (C) NC optimized conditions increase the yield of Sox10::GFP positive cells greater than 20-fold (53% ± 14%, n=4, p=0.002) over the DSi condition. (D) Using FACS analysis, Sox10::GFP was first detected at day 6 of NC differentiation and peaked by day 11. Sox10::GFP induction efficiency is represented as a percentage of total viable cells. (E) Significantly up- (green) and downregulated (red) genes at day 11 in NC-induced cells compared to DSi cells. Bold genes represent top 10 most differentially regulated genes. All error bars represent the s.e.m of at least three independent experiments. ** p<0.01. See also Figures S1 and S2.